Molecular characterization of “inverted” pericentromeric heterochromatin of chromosome 3

Summary Inversion of the pericentromeric region of human chromosome 3 [inv (3) (p11q11.2)] is a rare event. Initially, this inversion was identified with staining for Q-bands by fluorescence using quinacrine (QFQ) and later characterized with staining for C-bands by CBG technique. The molecular methods of fluorescence in situ hybridization (FISH) and AluI/Giemsa and TaqI/Giemsa techniques were utilized. The findings suggest that the variable band q11.2 on chromosome 3 contains alphoid DNA sequences, which appear to be similar to those identified by conventional methods in the centromeric region (band p11).     

Fatty-acid composition of polar lipids in fruit and leaf chloroplasts of “16:3”- and “18:3”-plant species

The fatty-acid composition of polar lipids from fruit and leaf chloroplasts was compared in five Solanaceous and two cucurbit species. The acylated fatty acids in monogalactosyl diglycerides (MGDG) from leaf chloroplasts of all five Solanaceous species included substantial amounts of ^7,10,13-hexadecatrienoic acid (16:3). In contrast, the MGDG from fruit chloroplasts of the Solanaceae contained very little of this plastid-specific polyunsaturate, and instead included a proportionately greater percentage of linoleic acid (18:2). In MGDG from leaf chloroplasts of two cucurbits, -linolenic acid (18:3) constituted 94–95% of the acylated fatty acids. Fruit-chloroplast galactolipids of the cucurbits had a greater abundance of 18:2, and hence a higher 18:2/18:3 ratio, than found in the corresponding leaf lipids. Among the phosphoglycerides, the unusual fatty acid 3-trans-hexadecenoate (trans-16:1) constituted from 15 to 24% of the acylated fatty acids in phosphatidyl glycerol (PG) from leaf chloroplasts (all species). In sharp contrast, trans-16:1 was virtually absent in PG from fruit chloroplasts of both Solanaceous and cucurbit species, and was replaced by a proportionate increase in the content of palmitate (16:0). The observed differences in the polar lipid fatty-acid composition of fruit and leaf chloroplasts are discussed in terms of the relative activity of several intrachloroplastic enzymes involved in lipid synthesis and fatty-acyl desaturation.Abbreviations MGDG monogalactosyldiglyceride - DGDG digalactosyl diglyceride - PC phosphatidyl choline - PE phosphatidyl ethanolamine - PG phosphatidyl glycerol     

A New Family of “H3L-Like” Histone Genes

The H3L histone variant gene in Paracentrotus lividus (sea urchin) shows almost all typical features of the replication-dependent histone genes, but it codes for the H3.3 histone protein with the S.//. A.IG amino acid motif, which is typical of the variants synthesized in a replication-independent manner. H3L-like histone genes have been found in several unrelated organisms. These genes are intronless and encode for the typical H3.3 histone proteins. The newly described family of H3L-like variants, nearly ubiquitous within the animal kingdom, could represent the common ancestor of H3 and H3.3 histone genes.     

Complement C3 polymorphism in sixteen populations of Colombia. Results of “Expedicion humana”

Blood samples of 1.022 individuals of both sexes, collected during the “Expedicion Humana” programme. Twelve Indian group, two Negroes, two urban and one mestizo populations of Colombia were C3 typed. The results showed the urban samples have C3*F genotype frequencies (17 and 20%) similar to the Spanish population. The Negroe system, with lower C3*F gene frequencies (3.5 and 7%) also resembled their parental populations in Africa. The Mestizo population reported in this study showed a lower value of the C3*F gene (2.2%) than previously studied. suggesting that the degree of admixture may vary in different regions of the country. As regards the Indian populations, there was a wide range of variation of the C3*F gene (0–19%) many tribes being monomorphic. This gene may be a marker of recent admixture in Amerindians, but there may be other genetic factors, such as selection and inbreeding in maintaining high levels of genetic polymorphism in tribes such as Guahibo.     

“Virtues” of apartheid

The traditional clinical method, which has served medicine well for over 100 years, had its origins in the integration of physical examination with morbid anatomy in early 19th-century France. Now this method is showing signs of failing to meet some contemporary needs. In particular, there is no means for understanding the inner experience of patients. Previous models of a transformed method have not grappled with the problem of validation. Data on the inner experience of patients are not open to empiric validation in the same way as clinical data. The process of understanding the meaning of an illness is not, therefore, scientific in the conventional sense. There are, none the less, rigorous methods for validating the results of this form of inquiry, notably those of phenomenology. A transformed method should aim to understand the meaning of an illness for the patient as well as provide a clinical diagnosis. The transformation will require a change in the epistemology of medicine and an educational process that encourages reflection and growth of self-knowledge.     

Structural Aspects and Chaperone Activity of Human HspB3: Role of the “C-Terminal Extension”

HspB3, an as yet uncharacterized sHsp, is present in muscle, brain, heart, and in fetal tissues. A point mutation correlates with the development of axonal motor neuropathy. We purified recombinant human HspB3. Circular dichroism studies indicate that it exhibits β-sheet structure. Gel filtration and sedimentation velocity experiments show that HspB3 exhibits polydisperse populations with predominantly trimeric species. HspB3 exhibits molecular chaperone-like activity in preventing the heat-induced aggregation of alcohol dehydrogenase (ADH). It exhibits moderate chaperone-like activity towards heat-induced aggregation of citrate synthase. However, it does not prevent the DTT-induced aggregation of insulin, indicating that it exhibits target protein-dependent molecular chaperone-like activity. Unlike other sHsps, it has a very short C-terminal extension. Fusion of the C-terminal extension of αB-crystallin results in altered tertiary and quaternary structure, and increase in polydispersity of the chimeric protein, HspB3αB-CT. The chimeric protein shows comparable chaperone-like activity towards heat-induced aggregation of ADH and citrate synthase. However, it shows enhanced activity towards DTT-induced aggregation of insulin. Our study, for the first time, provides the structural and chaperone functional characterization of HspB3 and also sheds light on the role of the C-terminal extension of sHsps.     

In situ hybridization of “Nick-translated” 3H-ribosomal DNA to chromosomes from salamanders

A technique is described for preparation of ³H-labelled DNA by nick-translation employing deoxyribonuclease I and DNA polymerase I. The labelled DNA can be obtained in high yield with specific activities of 10⁶ cpm/g or more. Ribosomal DNA, isolated from ovaries of young Xenopus laevis, and whole DNA from Plethodon cinereus were labelled in this way. The rDNA was used for in situ hybridization to meiotic chromosomes from P. cinereus, P. vehiculum and P. dunni. Autoradiographs of in situ hybrids were exposed for 5 to 10 days, by which time nucleolus organizer regions on the chromosomes of all 3 species were clearly and specifically labelled. In all eases, labelling was confined to a short region near the middle of the short arm of both halves of a medium length bivalent. It is concluded that nick-translation is a useful and altogether efficient method of labelling nucleic acids for subsequent use in experiments involving in situ hybridizations.     

“One-Pot” Syntheses of Malondialdehyde Adducts of Nucleosides

Short, “one-pot” syntheses of malondialdehyde adducts of deoxyguanosine, deoxyadenosine, and deoxycytidine are described. These syntheses proceed in improved yield and easier purification than previous syntheses and are well suited for the preparation of isotopically labeled nucleoside adducts for biomarker and metabolic studies.     

The amino acid sequence of “heavy chain disease” protein ZUC. Structure of the Fc fragment of immunoglobulin G3

The sequence of the Fc fragment of human IgG3 was studied, using a naturally-occurring γ3 heavy chain variant (ZUC). Though the molecule is internally deleted, it contains 248 residues, including the entire Fc fragment. The almost complete sequence of the C_H2 and C_H3 domains (position 234 to 446) indicates an extremely close evolutionary relationship with γ1 and γ4 chains. There is a 95% homology between IgG3 and IgGl and 92% between IgG3 and IgG4 in the C_H2 in the C_H3 domains.     

“Facilitation” and “Reversal of Response” of Neurones in the Cerebral Cortex

IN the course of experiments in which movements were elicited by direct electrical stimulation of the cerebral cortex, Graham-Brown^1–4 and Sherrington⁵ found that, after each train of stimuli, there were prolonged changes in the excitability of the cortex. They showed that the motor responses of forelimb muscles in primates, even though elicited by identical cortical stimuli, could be extremely variable. Two of the phenomena they described in detail (see also Lilly⁶) were “facilitation” (the increased motor response resulting from a second train of stimuli when it was applied within 20 s after the first train) and “reversal of response” (when repeated, identical stimulation of the same cortical point elicited flexion instead of extension, or vice versa).     

Dynamics of “neuron ring”

Summary Dynamics of neuron ring has been studied by means of computer simulation. The ring is formed of some number of identical neurons which loop together with lateral inhibitions and are stimulated by a command neuron. In this model, we assumed that each neuron has individual activity intrinsically without any inputs. The function of each neuron is essentially the same as that of Reiss' model; each neuron receives one excitatory and two inhibitory signals and when the membrane potential exceeds the instantaneous values of threshold, the neuron fires and sends out signals. Fatigue effect is also introduced in this model; after firing, the threshold level rises up for a period. This report is mainly concerned with the mechanism by which the individual activities of each neuron transit into an ordered behavior as a whole when they are externally stimulated. Results can be summarized as follows: The individual random activities of neurons can be organized and transfered into an ordered behavior on a completely symmetrical neuron ring if externally stimulated and the following conditions are satisfied; 1. the individual inherent activity is within a moderate range, 2. the fatigue effect is large enough, 3. external stimulation is high enough and 4. neurons in the ring are laterally inhibited. The individual activity and the lateral inhibition are essential to generate a meaningful output pattern as a group and fatigue effect acts as a stabilizing factor of the generated pattern.     

The Teaching of “Adaptation”

The use of the data from various athletic sports for exercises in the interpretation of data is proposed, and accounts are given of two such exercises. The first looks at the relationship between time and distance in world records for men running. When the logarithm of the time is plotted against the logarithm of the distance a straight line is produced. Thefactors that govern the speeds at which various distances are run are discussed and the application of these to other sports and to animals other than man is briefly mentioned. The second exercise looks at anomalous performances by men running in the 1968 Olympic Games which took place at high altitude. It is shown that while the lowered air resistance increased the speeds in sprint events, and the lowered oxygen content slowed down speeds in long distance events, these were not the only factors involved in limiting speeds.

Among the advantages of the use of such data are that they are readily and easily available, that they are the result of a lengthy selection process involving many subjects, that they appeal to students interested in sport and that they can link courses in biology and physical education.     

Flagellation of“Gluconobacter” melanogenus

Summary Electron-micrographs ofGluconobacter melanogenus strain AC 8 (formerlyG. liquefaciens), andG. melanogenus strain U 4, have conclusively confirmed the findings ofShimwell andCarr (1959),Stouthamer (1960), andLeifson (private communication), that these strains are not polarly flagellatedAcetomonas orGluconobacter strains, but peritrichously flagellated acetate-oxidisingAcetobacter ones. When transferred to the latter genus all evidence that these two genera are derived from a “common pool of ancestors”, as claimed byDe Ley (1961), disappears.     

Coleridge's “Theory of Life”

Coleridge has been seen by some not so much as a poet spoiled by philosophy, but as a philosopher who was also a poet. It could be argued that his major endeavor was an attempt to save the life sciences form the mechanistic interpretation which he saw as the outcome of Lockean “mechanico-corpuscularian” philosophy. This contribution describes that endeavour. It shows its connection to the social circumstances of the time. It discussess its relationship to the poetic sensibility of the “Lake poets” and to the German thought which Coleridge absorbed during and after his sojourn in Gottingen in 1798--99. It describes the nature of his “Theory of Life” as seen not only from the posthumous publication itself, but also from the numerous hints and struggles recorded in his voluminous notebooks, letters and lecture notes. It is concluded that, although never adequately assembled, it forms the only serious attempt to construct a profound alternative to the ultimately mechanistic biology of Charles Darwin and the physiologists of the second half of the century. As such it strongly influenced the young Richard Owen and, as is well known, was eventually overwhelmed by the Darwin-Huxley synthesis of the 1860s. Nevertheless, insofar as Coleridge's concept of life ultimately derived from his ambition to find a way of healing the Cartesian divide, we may wonder whether the recent upsurge in consciousness studies may cause us to look again at his panentheistic ideas and, discarding the obsolete and fanciful metaphysics, recast them into a more acceptable form. This revised version was published online in July 2006 with corrections to the Cover Date.     

“Mild” Uncoupling of Mitochondria

Recently, it was proposed that the thyroid hormone-mediated uncoupling in mitochondria is involved in the cellular defence system against free radicals (Skulachev V.P. (1996) Quart. Rev. Biophys. 29:169–202). This phenomenon was named mild uncoupling. It was postulated to be a protein-mediated process controlled by several factors. The data reported during the past 40 years, pointing to the protein-mediated uncoupling mechanism in mitochondria, are reviewed in a context of hypothetical properties of mild uncoupling. The mechanism of mild uncoupling is suggested to be the following: (a) mitochondria possess protein(s) that regulate the proton permeability of inner mitochondrial membrane; (b) these proteins are regulated by binding of an unidentified low-molecular-weight endogenous compound with properties resembling those of the most active artificial uncouplers like FCCP and SF6847; (c) the interaction of this compound with its target protein(s) is modulated by a thyroid hormone in a positive (i.e. enhancing the proton permeability) way and by sex steroid hormones in a negative way; (e) endogenous fatty acids can attenuate the influence of both thyroid and steroid hormones.     

“Maturation” of DNA duplexes

Reassociation of typical single-copy DNAs, like E. coli DNA, even when performed at relatively low temperatures, results in the formation of perfect duplexes with thermal stability very close to that of the native DNA. In contrast, duplexes of mouse repeated DNA as well as duplexes of Streptomyces DNA prepared under the same conditions, show a low thermal stability and undergo post-reassociation changes upon prolonged incubation. These changes, called maturation of the DNA duplexes, result in increasing of their thermal stability. Some of the factors affecting the rate of maturation are studied. The implication of the maturation process in reassociation analysis and in characterization of the heterogeneity of DNA is discussed.